Gao Mingxia, Deng Chunhui, Lin Shuang, Hu Fengli, Tang Jia, Yao Ning, Zhang Xiangmin
Department of Chemistry, Fudan University, Shanghai, China.
J Sep Sci. 2007 Apr;30(6):785-91. doi: 10.1002/jssc.200600372.
The most basic task in proteomics remains the detection and identification of proteins from a biological sample, and the most traditional way to achieve this goal consists in protein separations performed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Yet the 2-D PAGE-mass spectrometry (MS) approach has its drawbacks with regard to automation, sensitivity, and throughput. Consequently, considerable effort has been devoted to the development of non-gel-based proteome separation technologies in an effort to alleviate the shortcomings of 2-D PAGE. In addition, traditional Chinese medicines (TCMs), due to their long period of clinical testing and reliable therapeutic efficacy, are attracting increased global attention. However, hundreds or even thousands of components are usually present in TCMs, which results in great difficulties of separation. As a mainstream separation tool, multidimensional liquid separation systems have shown powerful separation ability, high peak capacity, and excellent detectability in the analysis of complex samples including biological samples and TCMs, etc. Therefore, this review emphasizes the most recent advances in multidimensional liquid chromatography and capillary electrophoresis-based separation techniques, and the corresponding applications in proteomics and TCMs. In view of the significant contributions from Chinese scientists, this review focuses mainly on the work of Chinese scientists in the above fields.
蛋白质组学中最基本的任务仍然是从生物样品中检测和鉴定蛋白质,而实现这一目标最传统的方法是通过二维聚丙烯酰胺凝胶电泳(2-D PAGE)进行蛋白质分离。然而,二维聚丙烯酰胺凝胶电泳-质谱联用(2-D PAGE-MS)方法在自动化、灵敏度和通量方面存在缺陷。因此,人们投入了大量精力来开发非基于凝胶的蛋白质组分离技术,以努力弥补二维聚丙烯酰胺凝胶电泳的不足。此外,中药因其长期的临床检验和可靠的治疗效果而受到全球越来越多的关注。然而,中药通常含有数百甚至数千种成分,这导致分离难度极大。作为主流的分离工具,多维液相分离系统在分析包括生物样品和中药等复杂样品时表现出强大的分离能力、高峰容量和出色的可检测性。因此,本综述着重介绍了基于多维液相色谱和毛细管电泳的分离技术的最新进展,以及它们在蛋白质组学和中药领域的相应应用。鉴于中国科学家的重大贡献,本综述主要聚焦于中国科学家在上述领域的工作。